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1、VIROLOGICASINICA2016,31(1):78–80DOI:10.1007/s12250-015-3703-3LETTERLongitudinalsurveillanceofSARS-likecoronavirusesinbatsbyquantitativereal-timePCRDearEditor,are?expressed?as?genome?copies?per?gram?of?bat?feces(copies/g).The?2002–2003?severe?acute?respiratory?syndromeThe?qRT-PCRs?were?perfo
2、rmed?using?an?AgPath-coronavirus?(SARS-CoV)?(Drosten?et?al.,?2003)?causedID??One-Step?RT-PCR?Kit?(Applied?Biosystems)?ac-human?pandemics?that?began?in?China?and?spread?glob-cording?to?the?manufacturer’s?instructions.?Each?25-μLally.?Subsequently,?diverse?SARS-like?coronavirusesreaction?mixt
3、ure?contained?12.5?μL?of?2×?RT-PCR?buf-(SL-CoVs)?have?been?detected?in?horseshoe?bats?infer,?1?μL?of?RT-PCR?Enzyme?Mix,?400?nmol/L?eachChina,?Europe,?and?Africa?(Li?et?al.,?2005;?Tong?et?al.,primer,?120?nmol/L?probe?primer,?and?1?μL?of?nucleic2009;?Drexler?et?al.,?2010).?Recently,?we?found?
4、SL-CoVsacid?extract.?Amplification?was?carried?out?in?96-wellwith?high?genetic?diversity?in?a?single?bat?colony?pre-plates?using?the?StepOne?PCR?system?(Applied?Biosys-dominantly?roosted?by?Chinese?horseshoe?bats?(Rhinolo-tems).?Thermocycling?conditions?were?as?follows:?10phussinicus)?in?Ku
5、nming,?Yunnan?province?(Ge?et?al.,min?at?45?°C?for?reverse?transcription,?10?min?at?95?°C2013).?Two?of?these?SL-CoVs?are?able?to?use?humanfor?activation?of?the?Taq?DNA?polymerase,?and?40?cyclesACE2?as?a?receptor?for?cell?entry?(Ge?et?al.,?2013;?Men-of?95?°C?for?15?sec?and?60?°C?for?45?sec.?
6、Each?run?in-achery?et?al.,?2015),?highlighting?the?risk?of?this?group?ofcluded?three?viral?positive?control?templates?and?one?neg-viruses?to?humans?and?the?importance?of?long-term?sur-ative?control?to?evaluate?assay?performance.?A?positiveveillance.result?was?defined?as?a?well-defined?expon
7、ential?fluores-We?conducted?a?longitudinal?surveillance?study?of?batcence?curve?that?crossed?the?cycle?threshold?(Ct)?withinSL-CoVs?using?quantitative?real-time?PCR?(qRT-PCR)38?cycles.?A?specimen?with?a?Ctvalue?>36?was?assayedtargeting?the?nucleocapsid?(
